Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: TNF-α modulates the WISP1 expression in stroma cells of the human bladder. (A) The RT-qPCR determined the expression of WISP1 in the human bladder cells. (B) Two WISP1 isoforms (WISP1v1 and WISP1v2) were found in the human bladder fibroblast (HBdSF) and bladder smooth muscle (HBdSMC) cells by RT-PCR with the pair of primers as shown. The expressions of WISP1, IL-6, CXCL5, CXCL12, and GDF15 of the HBdSF (C) and HBdSMC (D) cells after 10 ng/ml TNF-α treatment assessed by RT-qPCR. WISP1 (E) and IL-6 (F) secretions after TNF-α (0 – 20 ng/mL) treatments in HBdSF cells. Secretion of WISP1 (G) and IL-6 (F) in HBdSMC_shCOL and HBdSMC_shWISP1 cells treated with/without 10 ng/ml of TNF-α. Data are presented as the mean percentage (± SE; n = 4) in relation to the vehicle-treated HBdSMC_shCOL cells. (I) WISP1 secretion after 20 μM CAPE and/or TNF-α in HBdSF cells determined by ELISA assays. Data are presented as the mean percentage (±SE; n = 4) in relation to the vehicle-treated HBdSF cells. (J) The luciferase activity of the WISP1 reporter vector when HBdSMC cells were treated with TNF-α (10 ng/mL) and/or 20 μM CAPE for 24 h. * p < 0.05; ** p < 0.01; ND: no detectable.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of the conditioned media from the ectopic WISP1 overexpressed-293T cells on the cell proliferation in the bladder stroma cells. Expressions of WISP1v1 and WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the 293T cells were assessed by the RT-qPCR (A) and ELISA (B) assays. The ability of the proliferation of HBdSMC (C, D) and HBdSF (E, F) cells after being treated with the supernatant from 293T-DNA, 293T-WISP1v1, and 293T-WISP1v2 cells, respectively, was measured by the EdU staining or EdU flow cytometry assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. * p < 0.05; ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: WISP1 modulates expressions of α-SMA, IL-6, GDF15, and CXCL5 in bladder stroma cells. The mRNA levels of two WISP1 isoforms (WISP1v1 and WISP1v2) after WISP1 knock-downed in the HBdSF (A) and HBdSMC (B) cells were determined by RT-PCR. The protein levels with quantification analysis of WISP1, GDF15, and α-SMA of HBdSMC (C) and HBdSF (D) cells after mock-knockdown or WISP1-knockdown were assessed by immunoblot assays. The mRNA levels of the WISP1, α-SMA, IL-6, GDF15, and CXCL5 after knockdown of WISP1 in the HBdSMC (E) and HBdSF (F) cells, as indicated, were determined by RT-qPCR. Data are presented as target genes/β-actin of mock-knockdown cells relative to WISP1-knockdown cells. ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1-knockdown on the cell proliferation, migration, and contraction in bladder stroma and cancer cells. (A) The ability of cell proliferation of mock-knockdown HBdSMC (HBdSMC_shCOL) and WISPI-knockdown (HBdSMC_shWISP1) cells was assessed by EdU proliferation assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. (B) The ability of cell contraction of the mock-knockdown HBdSF (HBdSF_shCOL) and the WISP1-knockdown HBdSF (HBdSF_shWISP1) cells was measured by the collage contraction assays. Data are presented as gel area of the times as indicated in relation to the 0 h. (C) The ability of the migration of T24 cells after being treated with the supernatant from HBdSMC_shCOL and HBdSMC_shWISP1 cells, respectively, was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Knockdown, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1 on the cell proliferation and invasion in bladder cancer cells. (A) Cell proliferation of HT1376 and T24 cells after treatments of recombinant human WISP1 was assessed by CyQUANT cell proliferation assay. Data are presented as the mean percentage of the rhWISP1-treated cells relative to vehicle-treated cells. Expressions of total WISP1, WISP1v1, or WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the HT1376 cells were assessed by RT-qPCR (B), RT-PCR (C), and ELISA (D) assays. The abilities of the proliferation of HT-DNA, HT-WISP1v1 (E), and HT-WISP1v2 (F) cells were measured by Ki67 flow cytometry assays. Data are presented as the mean percentage of the Ki67-positive cells relative to HT-DNA cells. The invasion ability of HT1376 cells after ectopic overexpression of WISP1v1 (G) and WISP1v2 (H) was determined by in vitro Matrigel invasion assays (± SE; n = 4). Data are presented as the mean percentage of the invasion cells relative to HT-DNA cells. * p < 0.05; ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Recombinant, CyQUANT Assay, Proliferation Assay, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Over Expression, In Vitro

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Determination of the molecular mechanism of WISP1v1 on the cell invasion in the bladder cancer cells. (A) The protein levels of NDRG1, KAI1, and Maspin of HT-DNA and HT-WISP1v1 cells were determined by immunoblot assays. (B) The luciferase activity of the NDRG1 reporter vector was shown after transiently overexpressing various amounts of WISP1v1 expression vectors in HT1376 and T24 cells, as indicated. (C) The luciferase activity of KAI1 and Maspin reporter vectors was shown as indicated after transiently overexpressing various dosages of WISP1v1 expression vectors in HT1376 cells. (D) The luciferase activity of NDRG1, KAI1, and Maspin reporter vectors, respectively, after transiently overexpressed pcDNA, WISP1v1, and WISP1v2 expression vectors as indicated in HT1376 cells. (E) The mRNA levels of the E-cadherin, N-cadherin, Slug, Snail, and Vimentin were determined by RT-qPCR. Data are presented as target genes/β-actin of HT-WISP1v1 cells relative to HT-DNA cells. The mRNA levels of the WISP1 (F), NDRG1, KAI1, Maspin, and IL-6 (G) after transient overexpression of WISP1v1 in T24 cells were determined by RT-qPCR. Data are presented as target genes/β-actin of T24-WISP1v1 cells relative to T24-DNA cells. (H) The invasion ability of T24 cells after the ectopic overexpression of WISP1v1 was determined by in vitro Matrigel invasion assays (± SE; n = 4). The ability of the migration (I) and quantification analysis (I) of T24 cells after being treated with the supernatant from 293T-DNA and 293T-WISP1v1 cells was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. * p < 0.05, ** p < 0.01.

Article Snippet: To determine whether WISP1 is expressed in the human bladder stroma and cancer cells, we performed an RT-qPCR assay using the WISP1 probes (Hs04234730_m1; Applied Biosystems Inc.) to detect the transcript variants of WISP1.

Techniques: Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Over Expression, In Vitro, Migration

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Differential expression of WISP1 in normal and tumor tissue samples. (A) Expression of WISP1 in pan-cancer tissues and adjacent normal tissues via TIMER2.0. (B) Expression levels of WISP1 in ESCA from GEPIA2 (|Log2FC|> 1, P < 0.01, log scale: log2 (TPM + 1), Jitter Size: 0.4; T: Tumor, N: Normal). (C) Expression of WISP1 mRNA in the ESCC dataset. (D–G) Results of IHC and WB analyses using 12 paired ESCC tissues and adjacent control samples (T: Tumor, N: Normal; scale bars = 100μm). (H) ROC curves predicting the prognostic ability of high WISP1 expression for 1-year, 3-year, and 5-year patient survival. *p < 0.05; **p < 0.01; ***p < 0.001,;****P < 0.0001.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Quantitative Proteomics, Expressing, Control

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: The impact of WISP1 expression on the survival of ESCC patients. (A–C) Kaplan-Meier analysis of overall survival for high-expression versus low-expression groups in the datasets GSE53624 , GSE53625 , and TCGA. (D–F) Time-dependent ROC analysis for patients in the datasets.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Identification of DEGs in ESCC, functional enrichment analysis, and functional annotation of WISP1. (A) Volcano plot shows the DEGs from four datasets. (B) A Venn diagram shows genes that are differentially expressed across all four datasets. (C) GO and KEGG analyses reveal the potential biological mechanisms of DEGs in the four datasets. (D, E) GeneMANIA and STRING databases identify target proteins and genes associated with WISP1, followed by enrichment analysis of the functions of these proteins and genes. (F) Visualization of enrichment analysis results.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Functional Assay

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Tumor microenvironment and immune cell infiltration analysis in the GSE53624 dataset. (A) Differences in the abundance of infiltrating immune cells between high-expression and low-expression groups. (B–D) Differences in stromal scores, immune scores, and estimate scores between high-expression and low-expression groups. (E) Evaluation of the expression of immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3) between high-risk and low-risk groups. (F) Correlation between WISP1 expression and immune checkpoint molecules (CD274, PDCD1, TIGIT, CD276, CTLA4, LAG3). Blue indicates positive correlation, red indicates negative correlation, and the numbers inside the boxes represent the magnitude of the correlation. *p < 0.05; **p < 0.01; ***p < 0.001; ****P < 0.0001.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of hallmark pathways for esophageal squamous cell carcinoma (ESCC) patients stratified by WISP1 expression levels. (A–C) Enrichment patterns of associated pathways in the GSE53624 , GSE53625 , and TCGA datasets. The analyses were evaluated using a significance threshold corrected for the False Discovery Rate (FDR).

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Computational prediction of drug sensitivity for ESCC patients stratified by WISP1 expression levels. Using WISP1 expression grouping derived from the GSE53624 dataset, the “oncoPredict” R package was employed to evaluate the drug response spectrum. **** indicates statistical significance at p < 0.0001.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Expressing, Derivative Assay

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: Functional characterization of WISP1 in a subpopulation of fibroblasts. (A) Gene Set Enrichment Analysis (GSEA) shows enrichment results of differentially expressed genes between WISP1_Fib_Negative and WISP1_Fib_Positive clusters within the c5.all.v2024.1.Hs.symbols gene set; (B, C) display the enrichment results of DEGs in the h.all.v7.1.symbols gene set. (D) Proportional distribution of WISP1_Fib_Negative and WISP1_Fib_Positive clusters in normal esophageal tissues and ESCC samples. (E, F) Comparative analysis of intercellular communication signal intensity among fibroblast subpopulations. (G, H) Molecular stratification of fibroblasts using lineage-specific markers (DCN/Decorin, IGFBP6/Insulin Like Growth Factor Binding Protein 6, MFAP5/Microfibril Associated Protein 5, ACTA2/Actin Alpha 2 Smooth Muscle, TAGLN/Transgelin, CTHRC1/Collagen Triple Helix Repeat Containing 1)into NFs and CAFs subtypes. (I) Validation of CAF-specific markers (FAP, COL1A1, COL3A1, COL4A1, COL10A1, MMP1, MMP11, MMP14) and expression patterns of WISP1 in CAFs via single-cell RNA sequencing.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Functional Assay, Binding Assay, Biomarker Discovery, Expressing, RNA Sequencing

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: WISP1 regulates cancer-associated fibroblast function and extracellular matrix remodeling. (A, B) Immunofluorescence multiplex staining of α-smooth muscle actin (αSMA) and fibroblast activation protein (FAP) in paired cancer-associated fibroblasts (CAFs, tumor-derived) and normal fibroblasts (NFs, adjacent non-tumor tissues) (scale bars = 50μm). (C) Western blot (WB) analysis of WISP1, αSMA, and FAP expression in CAFs versus NFs. (D, E) Lentiviral short hairpin RNA (shRNA)-mediated WISP1 knockdown in CAFs, validated by quantitative reverse transcription PCR (qRT-PCR) and WB. (F–K) Functional characterization of CAF proliferation (CCK-8/EdU), migration, and invasion (Transwell) post-WISP1 silencing (scale bars = 50μm). (M) ELISA quantification of secreted WISP1 in supernatants from untransfected CAFs, CAFs-shVector, and CAFs-shWISP1 at 0 h, 24 h, and 48 h (N) Schematic of indirect co-culture system for CAF-ESCC interaction analysis. (L, P) WB assessment of extracellular matrix (ECM)-remodeling markers (COL1A1, MMP14) in WISP1-depleted CAFs and rescue via recombinant human WISP1 (rhWISP1). (O, Q) Transwell assay comparing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells co-cultured with untransfected CAFs, CAFs-shVector, or CAFs-shWISP1(scale bars = 50μm). (R, S) Transwell assay assessing the migration and invasion capacities of KYSE150 (left) and Eca109 (right) cells in the CAFs-shWISP1 co-culture system following rescue experiments with rhWISP1 supplementation (scale bar = 50μm).Data are presented as mean ± standard deviation from three independent experiments. Statistical significance was analyzed by two-tailed Student’s t-tests. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Immunofluorescence, Multiplex Assay, Staining, Activation Assay, Derivative Assay, Western Blot, Expressing, shRNA, Knockdown, Reverse Transcription, Quantitative RT-PCR, Functional Assay, CCK-8 Assay, Migration, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Recombinant, Transwell Assay, Cell Culture, Standard Deviation, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: WISP1 drives esophageal squamous cell carcinoma progression via modulation of cancer-associated fibroblasts and immune microenvironment

doi: 10.3389/fimmu.2025.1586790

Figure Lengend Snippet: WISP1 regulates ECM remodeling in CAFs through STAT3 signaling. (A, B) Representative phospho-kinase antibody array membrane comparing CAFs-shVector and CAFs-shWISP1. Red boxes highlight phosphorylated STAT3 (Y705) signals. (C) Dose-response curve of STAT3 inhibitor Stattic in CAFs. (D) Western blot analysis of phosphorylated STAT3 (Y705), total STAT3, and GAPDH (loading control) in: CAFs-shVector, CAFs-shWISP1, CAFs-shWISP1 + rhWISP1 (0.8 μg/mL), and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM). (E) Western blot analysis of COL1A1, MMP14, and GAPDH in CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) and CAFs-shWISP1 + rhWISP1 (0.8 μg/mL) + Stattic (7 μM).

Article Snippet: Primary fibroblasts were infected with shWISP1 lentivirus or treated with recombinant human WISP1 (rhWISP1) protein( NP_003873.1 , Sino Biological, China; endotoxin level <0.1 EU/μg, as determined by Limulus Amebocyte Lysate assay), followed by three washes with PBS.

Techniques: Ab Array, Membrane, Western Blot, Control

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: TNF-α modulates the WISP1 expression in stroma cells of the human bladder. (A) The RT-qPCR determined the expression of WISP1 in the human bladder cells. (B) Two WISP1 isoforms (WISP1v1 and WISP1v2) were found in the human bladder fibroblast (HBdSF) and bladder smooth muscle (HBdSMC) cells by RT-PCR with the pair of primers as shown. The expressions of WISP1, IL-6, CXCL5, CXCL12, and GDF15 of the HBdSF (C) and HBdSMC (D) cells after 10 ng/ml TNF-α treatment assessed by RT-qPCR. WISP1 (E) and IL-6 (F) secretions after TNF-α (0 – 20 ng/mL) treatments in HBdSF cells. Secretion of WISP1 (G) and IL-6 (F) in HBdSMC_shCOL and HBdSMC_shWISP1 cells treated with/without 10 ng/ml of TNF-α. Data are presented as the mean percentage (± SE; n = 4) in relation to the vehicle-treated HBdSMC_shCOL cells. (I) WISP1 secretion after 20 μM CAPE and/or TNF-α in HBdSF cells determined by ELISA assays. Data are presented as the mean percentage (±SE; n = 4) in relation to the vehicle-treated HBdSF cells. (J) The luciferase activity of the WISP1 reporter vector when HBdSMC cells were treated with TNF-α (10 ng/mL) and/or 20 μM CAPE for 24 h. * p < 0.05; ** p < 0.01; ND: no detectable.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of the conditioned media from the ectopic WISP1 overexpressed-293T cells on the cell proliferation in the bladder stroma cells. Expressions of WISP1v1 and WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the 293T cells were assessed by the RT-qPCR (A) and ELISA (B) assays. The ability of the proliferation of HBdSMC (C, D) and HBdSF (E, F) cells after being treated with the supernatant from 293T-DNA, 293T-WISP1v1, and 293T-WISP1v2 cells, respectively, was measured by the EdU staining or EdU flow cytometry assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. * p < 0.05; ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: WISP1 modulates expressions of α-SMA, IL-6, GDF15, and CXCL5 in bladder stroma cells. The mRNA levels of two WISP1 isoforms (WISP1v1 and WISP1v2) after WISP1 knock-downed in the HBdSF (A) and HBdSMC (B) cells were determined by RT-PCR. The protein levels with quantification analysis of WISP1, GDF15, and α-SMA of HBdSMC (C) and HBdSF (D) cells after mock-knockdown or WISP1-knockdown were assessed by immunoblot assays. The mRNA levels of the WISP1, α-SMA, IL-6, GDF15, and CXCL5 after knockdown of WISP1 in the HBdSMC (E) and HBdSF (F) cells, as indicated, were determined by RT-qPCR. Data are presented as target genes/β-actin of mock-knockdown cells relative to WISP1-knockdown cells. ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1-knockdown on the cell proliferation, migration, and contraction in bladder stroma and cancer cells. (A) The ability of cell proliferation of mock-knockdown HBdSMC (HBdSMC_shCOL) and WISPI-knockdown (HBdSMC_shWISP1) cells was assessed by EdU proliferation assays. The quantitative analysis is presented as the mean percentage (±SE; n = 3) of EdU-positive cells. (B) The ability of cell contraction of the mock-knockdown HBdSF (HBdSF_shCOL) and the WISP1-knockdown HBdSF (HBdSF_shWISP1) cells was measured by the collage contraction assays. Data are presented as gel area of the times as indicated in relation to the 0 h. (C) The ability of the migration of T24 cells after being treated with the supernatant from HBdSMC_shCOL and HBdSMC_shWISP1 cells, respectively, was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Knockdown, Migration

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Modulating effect of WISP1 on the cell proliferation and invasion in bladder cancer cells. (A) Cell proliferation of HT1376 and T24 cells after treatments of recombinant human WISP1 was assessed by CyQUANT cell proliferation assay. Data are presented as the mean percentage of the rhWISP1-treated cells relative to vehicle-treated cells. Expressions of total WISP1, WISP1v1, or WISP1v2 after the ectopic-overexpressed WISP1v1 and WISP1v2 in the HT1376 cells were assessed by RT-qPCR (B), RT-PCR (C), and ELISA (D) assays. The abilities of the proliferation of HT-DNA, HT-WISP1v1 (E), and HT-WISP1v2 (F) cells were measured by Ki67 flow cytometry assays. Data are presented as the mean percentage of the Ki67-positive cells relative to HT-DNA cells. The invasion ability of HT1376 cells after ectopic overexpression of WISP1v1 (G) and WISP1v2 (H) was determined by in vitro Matrigel invasion assays (± SE; n = 4). Data are presented as the mean percentage of the invasion cells relative to HT-DNA cells. * p < 0.05; ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Recombinant, CyQUANT Assay, Proliferation Assay, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Over Expression, In Vitro

Journal: Translational Oncology

Article Title: WISP1 is the stromal-secreting oncoprotein via paracrine downregulation of NDRG1, KAI1, and Maspin in human bladder cancer cells

doi: 10.1016/j.tranon.2026.102680

Figure Lengend Snippet: Determination of the molecular mechanism of WISP1v1 on the cell invasion in the bladder cancer cells. (A) The protein levels of NDRG1, KAI1, and Maspin of HT-DNA and HT-WISP1v1 cells were determined by immunoblot assays. (B) The luciferase activity of the NDRG1 reporter vector was shown after transiently overexpressing various amounts of WISP1v1 expression vectors in HT1376 and T24 cells, as indicated. (C) The luciferase activity of KAI1 and Maspin reporter vectors was shown as indicated after transiently overexpressing various dosages of WISP1v1 expression vectors in HT1376 cells. (D) The luciferase activity of NDRG1, KAI1, and Maspin reporter vectors, respectively, after transiently overexpressed pcDNA, WISP1v1, and WISP1v2 expression vectors as indicated in HT1376 cells. (E) The mRNA levels of the E-cadherin, N-cadherin, Slug, Snail, and Vimentin were determined by RT-qPCR. Data are presented as target genes/β-actin of HT-WISP1v1 cells relative to HT-DNA cells. The mRNA levels of the WISP1 (F), NDRG1, KAI1, Maspin, and IL-6 (G) after transient overexpression of WISP1v1 in T24 cells were determined by RT-qPCR. Data are presented as target genes/β-actin of T24-WISP1v1 cells relative to T24-DNA cells. (H) The invasion ability of T24 cells after the ectopic overexpression of WISP1v1 was determined by in vitro Matrigel invasion assays (± SE; n = 4). The ability of the migration (I) and quantification analysis (I) of T24 cells after being treated with the supernatant from 293T-DNA and 293T-WISP1v1 cells was determined by wound healing assays. The white line indicated the average of the leading cellular edges, and the wound area size was calculated by Image J. * p < 0.05, ** p < 0.01.

Article Snippet: TaqManTM gene expression master mix and polymerase chain reaction (PCR) FAM dye-labeled TaqMan MGB probes for human WISP1 (Hs04234730_m1 for total isoforms and Hs00180245 for WISP1v1), α-SMA (Hs00426835_g1), IL-6 (Hs00985639_m1), GDF15 (Hs00171132_m1), CXCL5 (Hs01099660_g1), SDF-1/CXCL12 (Hs03676656_m1), NDRG1 (Hs00608387_m1), KAI1 (Hs00356310_m1), Maspin (Hs00985283_m1), E-cadherin (Hs01023894_m1), N-cadherin (Hs00169953_m1), Snail (Hs00195591_m1), Slug (Hs00161904_m1), Vimentin (Hs00185584_m1), and β-actin (Hs01060665_g1) were purchased from Thermo Fisher Scientific Inc. (Vilnius, Lithuania).

Techniques: Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Over Expression, In Vitro, Migration